荧光PCR熔解曲线法对抗结核药物耐药性的诊断价值  

Evaluation of the diagnostic value of fluorescence PCR melting curve analysis in detecting drug resistance to tuberculosis drugs

作　　者：李浩 范大鹏 岳永宁 商学钗 李奂谕 周锦阳 刘立宾 蔡龙 LI Hao;FAN Dapeng;YUE Yongning;SHANG Xuechai;LI Huanyu;ZHOU Jingyang;LIU Libing;CAI Long(Hangzhou Chest Hospital Affiliated to School of Medicine of Zhejiang University,Hangzhou 310003,China)

机构地区：[1]浙江大学医学院附属杭州市胸科医院,浙江杭州310003

出　　处：《浙江实用医学》2023年第1期9-12,30,共5页Zhejiang Practical Medicine

基　　金：浙江省医药卫生科技计划项目(2023KY195)。

摘　　要：目的 评价荧光PCR熔解曲线法对抗结核药物利福平、异烟肼、氟喹诺酮、二线注射类药物(卡那霉素、阿米卡星、卷曲霉素)耐药性的诊断价值。方法 收集2018年1月~2019年5月在浙江大学医学院附属杭州市胸科医院收治的培养阳性且具有荧光PCR熔解曲线和BACTEC MGIT 960药敏结果的205例资料。以BACTEC MGIT960药敏结果为金标准,评价荧光PCR熔解曲线法对抗结核药物利福平、异烟肼、氟喹诺酮、二线注射类药物耐药性的诊断价值。结果 荧光PCR熔解曲线法敏感而BACTEC MGIT960药敏耐药的例数,在利福平、异烟肼、氟喹诺酮、二线注射类分别为1、9、1、1例。荧光PCR熔解曲线法耐药而BACTEC MGIT960药敏敏感的例数,在利福平、异烟肼、氟喹诺酮、二线注射类分别为2、1、2、0例。两种技术方法的总体符合率,在利福平、异烟肼、氟喹诺酮、二线注射类分别为98.54%、95.12%、98.54%、99.51%。荧光PCR熔解曲线法检测利福平耐药性的敏感性、特异性、阳性预测值、阴性预测值、Kappa值分别是96.96%、98.84%、94.12%、99.42%、0.946,对异烟肼耐药性的敏感性、特异性、阳性预测值、阴性预测值、Kappa值分别是80.43%、99.37%、97.37%、94.61%、0.851,对氟喹诺酮耐药性的敏感性、特异性、阳性预测值、阴性预测值、Kappa值分别是93.33%、98.94%、87.50%、99.47%、0.895,对二线注射类药物耐药性的敏感性、特异性、阳性预测值、阴性预测值、Kappa值分别是85.71%、100.00%、100.00%、99.50%、0.921。结论 荧光PCR熔解曲线法对利福平、异烟肼、氟喹诺酮、二线注射类药物的耐药性有较好的辅助诊断价值,适合临床结核病患者的早期耐药性筛查。Objective To evaluate the diagnostic value of fluorescence PCR melting curve analysis in detecting drug resistance to tuberculosis drugs such as rifampicin,isoniazid,fluoroquinolones,and second-line injectable drugs(kanamycin,amikacin,capreomycin).Methods A total of 205 patient data,collected from January 2018 to May 2019 at Hangzhou Chest Hospital affiliated to School of Medicine of Zhejiang University,were included in this study.These patients had positive cultures and both fluorescence PCR melting curve analysis and BACTEC MGIT 960 drug susceptibility test results available.The diagnostic value of fluorescence PCR melting curve analysis in detecting drug resistance to tuberculosis drugs,including rifampicin(RIF),isoniazid(INH),fluoroquinolones,and second-line injectable drugs,was evaluated using BACTEC MGIT 960 drug susceptibility results as the gold standard.Results The number of cases detected as drug resistant by fluorescence PCR melting curve analysis and drug sensitive by BACTEC MGIT 960 were 1 、9、1、and 1 for rifampicin,isoniazid,fluoroquinolones,and second-line injectable drugs,respectively.On the other hand,the number of cases detected as drug sensitive by fluorescence PCR melting curve analysis and drug resistant by BACTEC MGIT 960 were 2 、1、2 and 0 for rifampicin,isoniazid,fluoroquinolones,and second-line injectable drugs,respectively.The overall concordance rates between the two methods were 98.54%,95.12%,98.54%,and 99.51% for rifampicin,isoniazid,fluoroquinolones,and second-line injectable drugs,respectively.The sensitivity,specificity,positive predictive value,negative predictive value,and Kappa value of fluorescence PCR melting curve analysis in detecting rifampicin resistance were 96.96% 、98.84% 、94.12% 、99.42% and 0.946,respectively.For isoniazid resistance,the values were 80.43% 、99.37% 、97.37% 、94.61% and 0.851.For fluoroquinolones resistance,the values were 93.33% 、98.94% 、87.50% 、99.47% and 0.895.For second-line injectable drugs resistance,the values were 85.71% �

关 键 词：抗结核药物 耐药性 荧光PCR熔解曲线 BACTEC MGIT960药敏 

分 类 号：R978.3[医药卫生—药品]